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We narrowed to 1,106 results for: Ide

Showing: 181 - 200 of 1106 results

  1. Quick Guide to Working with Drosophila Part 1: Getting Started with Flies

    Type
    Blog Post
    ...deposition of the fertilized egg by its mother. Inside the pupal case, the larva will metamorphose into...light in coloration and can look very plump. On the side of their abdomen will be a dark spot. This is meconium...visible through their translucent bodies. This is the ideal stage to select for females because you can be certain...have an allele that causes the hairs on the dorsal side of its thorax to look like stubble (sb). For a better...search box in the upper right hand corner. They have video tutorials under the help tab like this one to get...pathogens and their hosts. It’s not always clear which side he roots for. You can try to convince him to use...

  2. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    ...the Dark Side." --Darth Vader in "Return of the Jedi" While Vader was referring to the evil side of a mystical...and rewarding. But, what if an FP fusion with a resident endoplasmic reticulum (ER) protein instead localizes...Previously, we have described a number of practical considerations that should be taken into account when deciding...must therefore determine your cloning strategy and decide whether you will create an N- or C-terminal fusion...needs. Equally important, before cloning you must consider whether and how well your FP will function in ...secreted into the extracellular milieu. Many of the resident proteins within these compartments undergo significant...post-translational modifications including glycosylation, disulfide bond formation, and proteolytic cleavage. FPs ...

  3. CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video]

    Type
    Blog Post
    .... Watch the following video to learn Daniel and Matthew's procedures. The video details how to transfect...Harvard Medical School. Addgene is proud to present a video reprint of the CRISPR article "Generation of Genomic...the Journal of Visualized Experiments (JOVE). The video publication by Stuart Orkin and Daniel Bauer's labs.... Biallelically disrupted cells can be easily identified. Even in the case of clones with monoallelic ...for biallelic deletion clones:    Like the video? The Orkin and Bauer lab's related paper describes... as their techniques for creating deletions, identifying these deletions, and finally characterization...

  4. SpyLigase Irreversibly Locks Peptides Together for Efficient Cell Capture

    Type
    Blog Post
    ...just join one peptide tag to another peptide tag. Addgene: What is the advantage of peptide tags for building... locking two peptide tags together. SpyLigase overcomes limitations in the use of peptide tags, which ...assemblies? Howarth: Peptide tags can be very generic; you should be able to use the same peptide tag for all ...thanks for the T-shirt! References: SpyLigase peptide-peptide ligation polymerizes affibodies to enhance ...itself together, to produce irreversible protein-peptide interactions. In a study published in Proceedings...Developing a new tool Addgene: Where did you get the idea to develop SpyLigase?  Howarth: It came from our...the Streptococcus pyogenes protein domain made it ideal for this purpose? What led you to it? Howarth: Our...

  5. From our Table to Yours: An Inside Look at Lunch at Addgene

    Type
    Blog Post
    ...four of us would eat together every day, sharing ideas for the company, but also talking about sports, ...Addgene. We’re a small non-profit, so we can only provide free food on special occasions. However, employees...and make time for special projects? We borrowed an idea from the software folks and started Hack Days. During...plasmid protocols for our website. Free lunch is provided for participants during the Hack Days, and we’...Lunches” With more employees, Addgene has naturally divided into smaller functional teams. Our Executive Director...make time for free food, and these lunches would provide a friendly setting for people in different labs...Who knows what could come out of it? Perhaps new ideas or collaborations, or at least a shared sense that...

  6. Three Key Considerations for Precise Neuronal Targeting Using AAV Technologies

    Type
    Blog Post
    ...practices and guidelines. It was wonderful to see the development of this practical guide by the workshop...networks, and specific neuronal populations to consider. To tease apart how different neurons impact a... discussions as a publicly accessible practical guide which was recently published in Frontiers in Neuroanatomy...experimental approach you choose, it’s essential to consider the technical limitations of each approach and...science.   Check out the precise neuronal targeting guide here! Additional resources on the Addgene blog ...retrograde AAVs Resources on Addgene.org Read our AAV guide Learn more about our viral service Find retrograde...

  7. HA Frankenbody, a New Imaging Tool to Visualize Single Molecules and Nascent Peptides

    Type
    Blog Post
    ...living systems, and often do not fold properly once inside cells.  Developing the HA frankenbody To combat...genetically encoded antibody based probe that detects the widely used HA epitope in diverse cellular environments...epitopes (Ewert et al., 2004). This scFv hybrid provides the probe structure and stability from the scaffold...translated quickly by the ribosome. This feature provides HA frankenbody the ability to bind nascent HA ...a versatile tool that can be used to visualize a wide range of protein dynamics in vivo. In the future...Resources on Addgene.org Read our fluorescent protein guide Find plasmids for subcellular localization Find ...

  8. Form Your Own Peer Mentoring Group: A How-To Guide for Scientists

    Type
    Blog Post
    ... organization, contact me! I can provide a step-by-step, how-to guide to get one started. Advantages ...communication skills. The leader also provides materials such as pre-reads, videos or exercises to be completed...increases the diversity of input and perspectives Provides a good mechanism for accountability that makes...to talk about?” There are an infinite number of ideas on the internet, but start by downloading my handy...collected. The next blog in this series will have more ideas on what to do once you are in a mentoring relationship...Peer Mentoring relationships? Comment below with ideas from your own mentoring group or with any questions...

  9. Download Addgene’s Science Career Guide, Plus 8 Career Tips from Addgenies

    Type
    Blog Post
    ...one comprehensive, downloadable guide: the Addgene Science Career Guide. We also asked Addgenies what their...endeavor.  Break the process down into steps and set aside time during the week to tackle each one.  Get the...you’d like to see discussed on the Addgene blog, consider writing a guest post. Additional resources on...careers blog posts Read our transferable skills guide Resources on Addgene.org Find job listings at ...

  10. Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation

    Type
    Blog Post
    ..., Moullier P, Ayuso E, Vandenberghe LH (2017) AAV-ID: A Rapid and Robust Assay for Batch-to-Batch Consistency...sequencing (NGS) QC platform that allows us to provide scientists with complete plasmid sequences. After... viral genome sequencing (VGS), to confirm the identity and serotype of our AAV preparations. We recently...an effective tool to confirm AAV vector genome identity. These studies used SMRT sequencing, TA-based ... that VGS can quickly and reliably confirm the identity of AAV genomes and detect contaminating AAV DNA...packaging cell line, helper and packaging plasmids. We decided to use this to our advantage by designing a Python...from multiple AAV lots can be run in parallel and provide serotype data in minutes and the scripts  can be...

  12. Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity

    Type
    Blog Post
    ...choice of junction sequences guided by experimentally-derived DNA Ligase fidelity data. Our work demonstrates...accuracy of assembly. The fidelity data can be applied to derive similar high-fidelity overhangs for any Golden...Reactions are often cycled between 37°C and 16°C to provide optimal reaction temps for both the restriction...appropriate levels of colony plating densities. Ligase fidelity and assembly efficiency Five fragment lacI/lacZ...redesigned for 12 and 24 fragments.  This redesign was guided by both advances in the re-engineering of the original...restriction enzyme and the completion of T4 DNA Ligase fidelity studies conducted by Potapov et al. at NEB (1,...Recently, NEB researchers profiled the comprehensive fidelity of cohesive end ligation by this enzyme for all...

  13. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

    Type
    Blog Post
    ...safe-targeting guides vs. non-targeting guides on screen analysis and hit calling. 5,644 non-targeting guides and...short guides had reduced toxicity compared to full-length guides. Reduced toxicity of short guides could...genes identified with short and long guides. Together these results suggests  that short guides have fewer...demonstrate safe-targeting guides as a potential control, and provide evidence that truncated gRNAs have...Genome-wide CRISPR/Cas9 screens are a high-throughput systematic approach for identifying genes involved...biological process. These screens provide an alternative to genome-wide RNAi screens, which although highly...questions about genome-wide CRISPR/Cas9 screen design Are non-targeting guides an appropriate control...

  16. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

    Type
    Blog Post
    ...This is the next step in high-throughput, genome-wide screening,” Konermann said. “If you want to activate...only do that screen with a system you are really confident about working for most genes and individual targeting...protein. SAM can be combined with a human genome-wide library to activate all 23,430 known coding isoforms... we could use information from the structure to guide and improve activator design,” she said. “Before...exposed RNA loops, which Konermann recognized as an ideal spot to fuse the activators instead of at the protein...consistency and validation with >80% effectively enriched guides per gene hit, and 100% validation of the top 10...genes of interest. They also “showed that different guide RNAs have different abilities to turn genes on and...

  17. CRISPR-Cas14: a family of small DNA-targeting enzymes enabling high-fidelity SNP genotyping

    Type
    Blog Post
    ...enzymes." Science 362.6416 (2018): 839-842. PubMed ID PMID: 30337455. Additional resources on the Addgene...systems were merely adaptive immune systems that provide bacteria protection against infectious agents. ...predicted that many others are still unknown. Identifying Cas14 variants In this quest to discover new ...that improved specificity of Cas14 enables high-fidelity SNP genotyping. The development of  Cas14-DETECTR...Find the Cas14 plasmids here Checkout our CRISPR guide page Browse the CRISPR collection ...

  19. A Needle in a Base-Stack: Cas9 Structural Biology

    Type
    Blog Post
    ...in the apo state (PDB ID 4CMP, top) compared to the guide RNA-bound state (PDB ID 4ZT0, bottom), seen from...structure (left, PDB ID 4ZT0) with Cas9:RNA:DNA electron microscopy structure (right, PDB ID 7S4X), viewed from...bound to guide RNA (PDB ID 4ZT0), shown opaque (left) and transparent (right). The RNA guide sequence ...Figure 5: S. pyogenes Cas9 bound to guide RNA and target DNA (PDB ID 7S4X). Residues R1333 and R1335 from...structure of S. pyogenes Cas9 bound to guide RNA and target DNA (PDB ID 7S4X), shown opaque (left) and transparent...Figure 7: S. pyogenes Cas9 bound to guide RNA and target DNA (PDB ID 7S4X). The end of the DNA:RNA heteroduplex...structure of S. pyogenes Cas9 in the apo state (PDB ID 4CMP) with domains colored as in Figure 1. HNH, red...

  20. More Data for You: Find Articles Citing Addgene Plasmids

    Type
    Blog Post
    ...with the Plasmid Name, Addgene Plasmid ID #, and the PubMed ID # (PMID) for the article that has used ...it has been validated in.  If a plasmid's Addgene ID # has been referenced in other publications, you'...used your plasmid of interest, which may help you identify relevant protocols and experimental data Find...
Showing: 181 - 200 of 1106 results